For collection of serum, peripheral blood was drawn by cardiac bleeding immediately after euthanasia. Blood samples were allowed to clot for at least 3 h at room temperature, and serum was separated following two rounds of centrifugation. Whole section of small intestine was homogenised in 2 ml RPMI (Life Technologies) and complete EDTA-free protease inhibitor (Roche) was added to inhibit protein degradation. Gut content and debris was removed by three rounds of centrifugation for collection of cytokine-containing supernatant. The concentration of IL-2, IL-4, IL-6, IL-10, IL-17A, IFN-γ and TNF was determined using the Mouse Th1/Th2/Th17 cytometric bead array (BD Bioscience) according to manufacturer’s instructions. Data was analysed using the FCAP Array software (BD Bioscience).
Cytometric bead array mouse th1 th2 th17
Manufactured by BD
Sourced in United States
The Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 is a multiplex assay that allows for the simultaneous detection and quantification of multiple cytokines in a single sample. The assay utilizes fluorescent-coded beads coated with analyte-specific capture antibodies to measure the concentration of cytokines associated with T-helper cell responses, including IFN-γ, IL-2, IL-4, IL-5, IL-6, IL-10, IL-17A, and TNF.
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Lab products found in correlation
Elisa, by R&D Systems (2 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions)
Complete edta free protease inhibitor, by Roche (1 mentions)
Fcap array software, by BD (1 mentions)
Meso scale discovery 35 u plex mouse biomarker group, by Mesoscale (1 mentions)
Meso scale quickplex sq 120, by Mesoscale (1 mentions)
Fcap array software 3, by BD (1 mentions)
Facscanto 2 flow cytometer, by BD (1 mentions)
Facscanto a, by BD (1 mentions)
7 protocols using cytometric bead array mouse th1 th2 th17
1
Cytokine Profiling of Mouse Serum and Intestine
2
T-cell Cytokine Profiling via CBA
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Extracellular secreted cytokine analysis was performed on cell media from T-lymphocytes cultured for 96 hours. Analysis was performed using the mouse Th1/Th2/Th17 cytometric bead array (BD Biosciences #560485, San Jose, CA) as per manufacturer’s instructions. Briefly, media was combined with antibody-coated fluorescent beads for 7 specific cytokines used to profile various T-lymphocyte subtypes. Beads were analyzed on a LSRII flow cytometer for quantification of specific cytokines.
3
Cytokine Secretion Assessment in T-Lymphocytes
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Secreted cytokines were assessed by two different methods. At 48 and 96 h post-activation, cell media from T-lymphocyte cultures was analyzed using the mouse Th1/Th2/Th17 cytometric bead array (BD Biosciences #560485, San Jose, CA) as per manufacturer's instructions. Analysis beads were analyzed on a LSRII flow cytometer for quantification of specific cytokines. At 96 h post-activation, cell media was also analyzed using a Meso Scale Discovery 35 U-Plex Mouse Biomarker Group (#K15083K-1, Rockville, MD). Samples were analyzed using a Meso Scale QuickPlex SQ 120, and normalized to cell number.
4
Adipocyte-SVF Co-culture Cytokine Analysis
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Adipocytes (3x105 cells/mL) and cells from SVF (5x106 cells/mL) were cultured in 5% CO2 incubator at 37°C for 24 hours in the absence or presence of PMA (0.4mg/mL) and Ionomycin (5mg/mL). After incubation, the supernatant was collected and stored at -70°C until use. Cytokine levels were assessed by Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 (BD Biosciences, USA) and adiponectin secreted by the culture of adipocytes was assessed by ELISA (R&D System, USA).
5
Cytokine Profiling by CBA
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Cytokines in the serum samples were measured with Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 (BD Biosciences) according to the manufacturer’s instructions. The data were collected on a FACSCANTO II flow cytometer (BD Bioscience) and analysed by FCAP Array Software 3.0 (BD Bioscience).
6
Th1/Th2/Th17 Cytokine Profiling
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Culture media was analyzed using Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 (BD Biosciences, Heidelberg, Germany) following manufacturer's instructions. The measurement was performed in a FACSCanto A (BD Biosciences, Heidelberg, Germany) and data was analyzed using the Kitprovided Software (FCAP Array v3.0.1).
7
Cytokine Analysis of Adipocyte and SVF Cells
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Adipocytes (3x10 5 cells/mL) and cells from SVF (5x10 6 cells/mL) were cultured in 5% CO 2 incubator at 37°C for 24 hours in the absence or presence of PMA (0.4mg/mL) and Ionomycin (5mg/mL). After incubation, the supernatant was collected and stored at -70°C until use. Cytokine levels were assessed by Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 (BD Biosciences, USA) and adiponectin secreted by the culture of adipocytes was assessed by ELISA (R&D System, USA).
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